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Antarctic ice alga Chlamydomonas sp. ICE-L can endure extreme low temperature and high salinity stress under freezing conditions. To elucidate the molecular
acclimation mechanisms using gene expression analysis, the expression stabilities of ten housekeeping genes of Chlamydomonas sp. ICE-L during freezing stress were analyzed. Some discrepancies were detected in the ranking of the candidate reference
genes between geNorm and NormFinder programs, but there was substantial agreement between the groups of genes with the most
and the least stable expression. RPL19 was ranked as the best candidate reference genes. Pairwise variation (V) analysis indicated the combination of two reference genes was sufficient for qRT-PCR data normalization under the experimental
conditions. Considering the co-regulation between RPL19 and RPL32 (the most stable gene pairs given by geNorm program), we
propose that the mean data rendered by RPL19 and GAPDH (the most stable gene pairs given by NormFinder program) be used to
normalize gene expression values in Chlamydomonas sp. ICE-L more accurately. The example of FAD3 gene expression calculation demonstrated the importance of selecting an appropriate
category and number of reference genes to achieve an accurate and reliable normalization of gene expression during freeze
acclimation in Chlamydomonas sp. ICE-L. 相似文献
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Differential gene expression in nasopharyngeal carcinoma cells 总被引:15,自引:0,他引:15
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基因表达转录分析中内参基因的选择 总被引:17,自引:0,他引:17
目前基因表达的转录分析多采用单一看家基因作为内参来校正目标基因的表达量.实验中以人肝癌BEL-7402细胞为研究对象,应用实时荧光定量PCR技术,观察了新型三肽化合物酪丝缬肽作用后RPL13A、UBC、EIF4A、B2M、GAPDH和ACTB共6个看家基因mRNA水平的表达情况.经过geNorm程序统计学分析处理,结果表明,这6个看家基因的表达存在差异,确定了RPL13A、UBC2个看家基因用于校正目标基因的表达量.基因表达转录分析中内参基因选择的必要性在实验中得以证明,更重要的是为各种实验因素影响下(尤其是新物质作用下)内参基因的选择介绍和提供了一种行之有效的方法. 相似文献
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